Pacheco A B, Guth B E, Soares K C, Nishimura L, de Almeida D F, Ferreira L C
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Brazil.
J Clin Microbiol. 1997 Jun;35(6):1521-5. doi: 10.1128/jcm.35.6.1521-1525.1997.
The genetic diversity of 47 enterotoxigenic Escherichia coli (ETEC) strains of serotypes O6:H16, O27:H7, O29:H21, O128ac:H12, and O153:H45, previously isolated from diarrheic patients in Brazil over a period of 15 years, was investigated by random amplification of polymorphic DNA (RAPD). Informative band arrays were obtained with three 10-mer primers with G+C contents of 50, 60, and 70%. Based on the combination of the band profiles generated by the three primers 22 RAPD types were detected, and 5 major clonal clusters, each one with at least 80% identical bands, were established. The clonal clusters corresponded to strains having the same serotype which, in most cases, also had the same virulence factors (colonization factors and toxin types) and outer membrane protein and lipopolysaccharide sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles. The results suggested a correlation between phenotypic properties and genetic relatedness of ETEC isolates of human origin and indicated that a reduced number of clonally related strains are found in areas of ETEC endemicity in Brazil. Moreover, the RAPD technique revealed intraserotype-specific variations, undetectable by the combination of several phenotypic typing methods, among the ETEC strains analyzed. These results show that RAPD typing represents a useful tool for population genetics as well as for epidemiological studies of ETEC.
通过随机扩增多态性DNA(RAPD)技术,对先前在15年期间从巴西腹泻患者中分离出的47株血清型为O6:H16、O27:H7、O29:H21、O128ac:H12和O153:H45的产肠毒素大肠杆菌(ETEC)菌株的遗传多样性进行了研究。使用三种G+C含量分别为50%、60%和70%的10聚体引物获得了信息丰富的条带阵列。基于三种引物产生的条带图谱组合,检测到22种RAPD类型,并建立了5个主要克隆簇,每个克隆簇至少有80%的相同条带。这些克隆簇对应于具有相同血清型的菌株,在大多数情况下,它们还具有相同的毒力因子(定植因子和毒素类型)以及外膜蛋白和脂多糖十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图谱。结果表明,人类来源的ETEC分离株的表型特性与遗传相关性之间存在关联,并表明在巴西ETEC地方性流行地区发现的克隆相关菌株数量减少。此外,RAPD技术揭示了在所分析的ETEC菌株中存在几种表型分型方法组合无法检测到的血清型内特异性变异。这些结果表明,RAPD分型是ETEC群体遗传学以及流行病学研究的有用工具。